Use of an fgf-21 compound and a glp-1 compound for the treatment of obesity

ABSTRACT

The present invention provides methods of lowering body weight by administering an FGF-21 compound in combination with a GLP- 1  compound. In addition, the present invention also provides methods to treat obesity by administering an FGF- 21  compound in combination with a GLP- 1  compound. The present invention also discloses combinations useful in the methods of the present invention.

The present invention relates to compositions that comprise an FGF-21compound and a GLP-1 compound. These compositions can be used to lowerbody weight and treat obesity

BACKGROUND OF THE INVENTION

Obesity, and especially upper body obesity, is the most commonnutritional disorder in the over-nourished populations of the world.Current methods for promoting weight loss are not completelysatisfactory. Unfortunately, an estimated 33 billion dollars a year arespent on weight-loss measures that are largely futile. Thus, new methodsand compositions such as pharmaceutical agents that promote weight-lossare urgently needed to complement old approaches.

Fibroblast growth factor 21 (FGF-21) belongs to a family of largepolypeptides widely expressed in developing and adult tissues that playcrucial roles in multiple physiological functions. FGF-21 has beenreported to stimulate glucose-uptake in mouse adipocytes after prolongedtreatment, in the presence and absence of insulin, and to decrease fedand fasting blood glucose, triglycerides, and glucagon levels in ob/oband db/db mice in a dose-dependent manner, thus, providing the basis forthe use of FGF-21 as a therapy for treating diabetes and obesity(WO03/011213).

In addition to its beneficial effects on Type 2 diabetes, Glucagon-likepeptide-1 (GLP-1) compounds have been described for the treatment ofobesity. (WO98/019698). Although both FGF-21 and GLP-1 compounds haveshown positive effects in treating obesity, there has not been anyindication that a combination of FGF-21 compounds and GLP-1 compoundswould provide a synergistic effect on lowering body weight. There isthus, still a need for additional beneficial therapeutics for weightloss.

Applicants have determined that a combination of an FGF-21 compound anda GLP-1 compound have an unexpected synergistic effect on lowering bodyweight.

SUMMARY OF THE INVENTION

The present invention relates to compositions comprising a FGF-21compound and a GLP-1 compound. The present invention also provides amethod of lowering body weight comprising administering an FGF-21compound in combination with a GLP-1 compound. In another embodiment,the present invention provides a method of treating obesity comprisingadministering a FGF-21 compound in combination with a GLP-1 compound.

The present invention also provides compositions comprising a FGF-21compound and an exendin compound. The present invention also provides amethod of lowering body weight comprising administering an FGF-21compound in combination with an exendin compound. In another embodiment,the present invention provides a method of treating obesity comprisingadministering a FGF-21 compound in combination with an exendin compound.

DETAILED DESCRIPTION

A “FGF-21 compound” is defined as a compound comprising native humanFGF-21 (SEQ ID NO: 2), an FGF-21 analog, or an FGF-21 derivative. TheFGF-21 compounds of the present invention retain FGF-21 activity asmeasured in assays as described in Kharitonenkov, et al., (Journal ofClinical Investigation. 115(6):1627 (2005)).

A “FGF-21 analog” is defined as a molecule having a modificationincluding one or more amino acid substitutions, deletions, inversions oradditions when compared with SEQ ID NO: 2.

A “FGF-21 derivative” is defined as a molecule having the amino acidsequence of human FGF-21 (SEQ ID NO: 2) or of a FGF-21 analog butadditionally having at least one chemical modification of one or more ofits amino acid side groups, α-carbon atoms, terminal amino group, orterminal carboxylic acid group. Modifications at amino acid side groupsinclude acylation of lysine e-amino groups, N-alkylation of arginine,histidine, or lysine, alkylation of glutamic or aspartic carboxylic acidgroups, and deamidation of glutamine or asparagine. Modifications of theterminal amino include the des-amino, N-lower alkyl, N-di-lower alkyl,and N-acyl modifications. Modifications of the terminal carboxy groupinclude the amide, lower alkyl amide, dialkyl amide, and lower alkylester modifications. A lower alkyl is a C₁-C₄alkyl. Furthermore, one ormore side groups, or terminal groups, may be protected by protectivegroups known to the ordinarily-skilled protein chemist. The α-carbon ofan amino acid may be mono- or di-methylated. The chemical modificationmay also include “pegylation.”

Specific substitutions to human FGF-21, FGF-21 analog or FGF-21derivative indicated using the specific amino acid present aftersubstitution or modification at a particular residue followed by theresidue number. For example, Cys¹¹⁸-FGF-21 indicates that there has beena cysteine introduced at position 118 of human FGF-21.

A “GLP-1 compound” is defined as a compound comprising the amino acidsequence of native human GLP-1 (SEQ ID NO: 3), a GLP-1 analog or GLP-1derivative, which maintains GLP-1 activity. GLP-1 activity may bemeasured by methods known in the art, including using in vivoexperiments and in vitro assays that measure GLP-1 receptor bindingactivity or receptor activation, e.g., assays employing pancreatic isletcells or insulinoma cells, as described in EP 619,322, Gelfand, et al.and U.S. Pat. No. 5,120,712, respectively. GLP-1 compounds are wellknown in the art. See, e.g. PCT International Application PublicationNos. WO 03/040309, U.S. Pat. Nos. 6,593,295, 7,141,547, and 7,176,278.

A “GLP-1 analog” is defined as a molecule having a modificationincluding one or more amino acid substitutions, deletions, inversions,or additions when compared with SEQ ID NO: 3. “GLP-1 analog” alsoincludes “GLP-1 fusion proteins” where the GLP-1 fusion protein is aheterologous protein comprising a GLP-1 or GLP-1 analog and a secondpolypeptide selected from the group consisting of human albumin, humanalbumin analogs, fragments of human albumin, transferrin, transferrinanalogs, transferrin derivatives, fragments of transferring, the Fcportion of an immunoglobulin, an analog of the Fc portion of animmunoglobulin, and fragments of the Fc portion of an immunoglobulin,and wherein the C-terminus of the first polypeptide is fused to theN-terminus of the second polypeptide. The GLP-1 or GLP-1 analog may befused to the second polypeptide via a peptide linker. The GLP-1 fusionproteins of the present invention contain an Fc portion which is derivedfrom human IgG4 but comprises one or more substitutions compared to thewild-type human sequence (SEQ ID NO: 5).

A “GLP-1 derivative” is defined as a molecule having the amino acidsequence of native human GLP-1 or of a GLP-1 analog, but additionallyhaving at least one chemical modification of one or more of its aminoacid side groups, a-carbon atoms, terminal amino group, or terminalcarboxylic acid group. Modifications at amino acid side groups includeacylation of lysine e-amino groups, N-alkylation of arginine, histidine,or lysine, alkylation of glutamic or aspartic carboxylic acid groups,and deamidation of glutamine or asparagine. Modifications of theterminal amino include the des-amino, N-lower alkyl, N-di-lower alkyl,and N-acyl modifications. Modifications of the terminal carboxy groupinclude the amide, lower alkyl amide, dialkyl amide, and lower alkylester modifications. A lower alkyl is a C₁-C₄alkyl. Furthermore, one ormore side groups, or terminal groups, may be protected by protectivegroups known to the ordinarily-skilled protein chemist. The α-carbon ofan amino acid may be mono- or di-methylated. The chemical modificationmay also include the pegylation of an amino acid of the peptide orpolypeptide.

The nomenclature used herein to refer to specific GLP-1 analogs andGLP-1 derivatives is defined as follows: Specific substitutions to aGLP-1 analog and GLP-1 derivative are indicated using the specific aminoacid being substituted followed by the residue number. GLP-1 (7-37)indicates that the GLP-1 analog portion of the mature fusion proteinbegins with His at position 7 and ends with Gly at position 37. In thecase of additions of amino acids to a GLP-1 amino acid sequence, theadded amino acid is indicated followed by the position at which it ispresent. For example, the addition of two serine residues to theC-terminus of wild type GLP-1 will be referred to as Ser³⁸-Ser³⁹-GLP-1.

An “exendin compound” is defined as a compound comprising the amino acidsequence of exendin-4 (SEQ ID NO: 4), an exendin-4 analog or exendin-4derivative, wherein the exendin compound maintains exendin-4 activity.

An “exendin-4 analog” is defined as a compound having a modificationincluding one or more amino acid substitutions, deletions, inversions,or additions when compared with the amino acid sequence of exendin-4(SEQ ID NO: 4). “Exendin-4 analog” also includes “exendin fusionproteins” where the “exendin fusion protein” is a heterologous proteincomprising an exendin-4 or exendin-4 analog and a second polypeptideselected from the group consisting of human albumin, human albuminanalogs, fragments of human albumin, transferrin, transferrin analogs,transferrin derivatives, fragments of transferring, the Fc portion of animmunoglobulin, an analog of the Fc portion of an immunoglobulin, andfragments of the Fc portion of an immunoglobulin, and wherein theC-terminus of the first polypeptide is fused to the N-terminus of thesecond polypeptide. The exendin-4 or exendin-4 analog may be fused tothe second polypeptide via a peptide linker. The exendin-4 fusionproteins of the present invention may contain an Fc portion which isderived from human IgG4 but comprises one or more substitutions comparedto the wild-type human sequence (SEQ ID NO: 5).

An “exendin-4 derivative” is defined as a compound having the amino acidsequence of exendin-4 or of an exendin-4 analog, but additionally havingat least one chemical modification of one or more of its amino acid sidegroups, a-carbon atoms, terminal amino group, or terminal carboxylicacid group. Modifications at amino acid side groups include acylation oflysine e-amino groups, N-alkylation of arginine, histidine, or lysine,alkylation of glulamic or aspartic carboxylic acid groups, anddeamidation of glutamine or asparagine. Modifications of the terminalamino include the des-amino, N-lower alkyl, N-di-lower alkyl, and N-acylmodifications. Modifications of the terminal carboxy group include theamide, lower alkyl amide, dialkyl amide, and lower alkyl estermodifications. A lower alkyl is a C₁-C₄alkyl. Furthermore, one or moreside groups, or terminal groups, may be protected by protective groupsknown to the ordinarily-skilled protein chemist. The α-carbon of anamino acid may be mono- or di-methylated. The chemical modification mayalso include pegylation.

As used herein, the Fc portion of an immunoglobulin has the meaningcommonly given to the term in the field of immunology. Specifically,this term refers to an antibody fragment which does not contain the twoantigen binding regions (the Fab fragments) from the antibody. The Fcportion consists of the constant region of an antibody from both heavychains, which associate through non-covalent interactions and disulfidebonds. The Fc portion can include the hinge regions and extend throughthe CH2 and CH3 domains to the c-terminus of the antibody. The Fcportion can further include one or more glycosylation sites.

The fusion proteins described herein may also contain a linker (“L”).The linker may comprise the sequenceGly-Gly-Gly-Gly-Ser-Gly-Gly-Gly-Gly-Ser-Gly-Gly-Gly-Gly-Ser (SEQ ID NO:6). The number immediately preceding the L refers to the number oflinkers separating the particular peptide or protein portion from the Fcportion. A linker specified as 1.5L refers to the sequenceGly-Ser-Gly-Gly-Gly-Gly-Ser-Gly-Gly-Gly-Gly-Ser-Gly-Gly-Gly-Gly-Ser-Gly-Gly-Gly-Gly-Ser(SEQ ID NO: 7). IgG4 refers to an analog of the human IgG4 Fc sequencespecified as SEQ ID NO: 5. Substitutions in the IgG4 Fc portion of thefusion protein are indicated in parenthesis. The wild-type amino acid isspecified by its common abbreviation followed by the position number inthe context of the entire IgG4 sequence using the EU numbering systemfollowed by the amino acid being substituted at that position specifiedby its common abbreviation.

“Pegylation” or “Pegylated” refers to a compound of the presentinvention that is chemically modified by covalent attachment of amolecule or molecules of polyethylene glycol or a derivative thereof.Furthermore, it is intended that the term “PEG” refers to polyethyleneglycol or a derivative thereof as are known in the art. In its typicalform, PEG is a linear polymer with terminal hydroxyl groups and has theformula HO—CH₂CH₂—(CH₂CH₂O)_(n)—CH₂CH₂—OH, where n is from about 8 toabout 4000. The terminal hydrogen may be substituted with a protectivegroup such as an alkyl or aryl group. Preferably, PEG has at least onehydroxy group, more preferably it is a terminal hydroxy group. It isthis hydroxy group which is preferably activated to react with thepeptide. There are many forms of PEG useful for the present invention.Numerous derivatives of PEG exist in the art and are suitable for use inthe invention. The PEG molecule covalently attached to compounds in thepresent invention is not intended to be limited to a particular type.PEG's molecular weight is preferably from 500-100,000 daltons and morepreferably from 20,000-60,000 daltons and most preferably from20.000-40,000 daltons. PEG may be linear or branched.

“In combination with” or “coadministration” refers to the administrationof a FGF-21 compound with a GLP-1 compound either simultaneously,sequentially or a combination thereof. The combination therapy of aFGF-21 compound with a GLP-1 compound results in a synergistic effect inlowering body weight and thus, in the treatment of obesity. Thecombination therapy also results in a synergistic effect on lowerelevated blood glucose levels and thus, a potential use in the treatmentof diabetes.

The term “synergy” or “synergistic” as used herein is defined to mean acombination of components wherein the effect of the combination isgreater than the additive individual effects of each component of thecombination. For purposes of the present invention, “effect” refers to aloss in body weight, lowering of body weight or a reduction in bloodglucose levels.

The FGF-21 compounds, the GLP-1 compounds and the exendin compounds ofthe present invention may be made using various techniques known to oneof skill in the art. For example, the FGF-21, GLP-1 and exendincompounds may be made using recombinant techniques. As the DNA sequencesof mature human FGF-21 (SEQ ID NO: 1), native GLP-1, and exendin-4 areknown, PCR methodology may be used to isolate genes encoding thecorresponding gene. Further, one of skill in the art is aware of variousmethods to introduce changes in the DNA sequence so as to effect desiredchanges in the amino acid sequence of the resulting FGF-21, GLP-1 orexendin compounds.

The GLP-1 and the exendin peptides of the present invention can also beprepared by using standard methods of solid-phase peptide synthesistechniques. Peptide synthesizers are commercially available from, forexample, Applied Biosystems in Foster City, Calif. Reagents for solidphase synthesis are commercially available, for example, from MidwestBiotech (Fishers, Ind.). Solid phase peptide synthesizers can be usedaccording to manufacturer's instructions for blocking interferinggroups, protecting the amino acid to be reacted, coupling, decoupling,and capping of unreacted amino acids.

A wide variety of methods have been described in the art to covalentlyconjugate PEGs to peptides (for review article see, Roberts, M. et al.Advanced Drug Delivery Reviews, 54:459-476, 2002). Pegylation ofpeptides at the carboxy-terminus may be performed via enzymatic couplingusing recombinant compounds of the present invention as a precursor oralternative methods known in the art and described. See e.g. U.S. Pat.No. 4,343,898 or International Journal of Peptide & Protein Research.43: 127-38, 1994.

Various methods of protein purification may be employed and such methodsare known in the art and described, for example, in Deutscher, Methodsin Enzymology 182: 83-9 (1990) and Scopes, Protein Purification:Principles and Practice, Springer-Verlag, NY (1982). The purificationstep(s) selected will depend on the nature of the production processused and the particular protein produced. For example, fusion proteinscomprising an Fc fragment can be effectively purified using a Protein Aor Protein G affinity matrix. Low or high pH buffers can be used toelute the fusion protein from the affinity matrix Mild elutionconditions will aid in preventing irreversible denaturation of thefusion protein.

Coadministration of FGF-21 compounds. GLP-1 compounds and exendincompounds of the present invention may be via any route known to beeffective by the physician of ordinary skill. Peripheral parenteral isone such method. Parenteral administration is commonly understood in themedical literature as the injection of a dosage form into the body by asterile syringe or some other mechanical device such as an infusionpump. Peripheral parenteral routes can include intravenous,intramuscular, subcutaneous, and intraperitoneal routes ofadministration.

Those skilled in the art can readily optimize pharmaceutically effectivedosages and administration regimens for the combinations of the presentinvention, as determined by good medical practice and the clinicalcondition of the individual patient. For example, a typical dose rangefor the FGF-21 compounds of the present invention will range from about0.01 mg per day to about 1000 mg per day for an adult. A typical doserange for the GLP-1 derivative compounds of the present invention willrange from about 0.01 mg per day to about 1000 mg per day for an adult.For GLP-1 fusion proteins, doses may be in the range of 0.01 to 1 mg/kgbody weight, preferably in the range of 0.05 to 0.5 mg/kg body weight.

In an embodiment, the present invention provides a compositioncomprising a FGF-21 compound and a GLP-1 compound. In anotherembodiment, the present invention provides a composition comprising aFGF-21 compound and a GLP-1 compound, wherein the FGF-21 compound isselected from the group consisting of human FGF-21, an FGF-21 analog andan FGF-21 derivative and wherein the GLP-1 compound is selected from thegroup consisting of GLP-1 analog, GLP-1 derivative and GLP-1 fusionproteins. In a preferred embodiment, the composition comprises an FGF-21analog and a GLP-1 analog. In another preferred embodiment, thecomposition comprises a FGF-21 analog and a GLP-1 derivative. In anotherpreferred embodiment, the composition comprises a FGF-21 analog and aGLP-1 fusion protein.

In another embodiment, the present invention provides a compositioncomprising an FGF-21 compound and an exendin compound. In a preferredembodiment, the composition comprises a FGF-21 compound and an exendincompound, wherein the FGF-21 compound is selected from the groupconsisting of human FGF-21, an FGF-21 analog and an FGF-21 derivativeand wherein the exendin compound is selected from the group consistingof exendin-4, an exendin-4 analog, an exendin-4 derivative, and anexendin-4 agonist. In a preferred embodiment, the composition comprisesa FGF-21 analog and exendin-4. In another preferred embodiment, thecomposition comprises a FGF-21 analog and an exendin-4 analog. Inanother preferred embodiment, the composition comprises a FGF-21 analogand an exendin-4 derivative. In another preferred embodiment, thecomposition comprises a FGF-21 analog and an exendin-4 agonist.

The present invention also provides a method of lowering body weightcomprising administering a FGF-21 compound in combination with a GLP-1compound. In a more preferred embodiment, administering a FGF-21compound in combination with a GLP-1 compound results in a synergisticeffect on weight loss. In another embodiment, the method of loweringbody weight comprises administering a FGF-21 compound in combinationwith a GLP-1 compound, wherein the FGF-21 compound is selected from thegroup consisting of human FGF-21, an FGF-21 analog and an FGF-21derivative and wherein the GLP-1 compound is selected from the groupconsisting of GLP-1 analog, GLP-1 derivative and GLP-1 fusion protein.In a preferred embodiment, the method of lowering body weight comprisesadministering a FGF-21 analog in combination with a GLP-1 analog. Inanother preferred embodiment, the method of lowering body weightcomprises administering a FGF-21 analog in combination with a GLP-1derivative. In another preferred embodiment, the method of lowering bodyweight comprises administering a FGF-21 analog and a GLP-1 fusionprotein.

The present invention also provides a method of lowering body weightcomprising administering a FGF-21 compound in combination with anexendin compound. In more preferred embodiment, administering a FGF-21compound in combination with an exendin compound results in asynergistic effect on weight loss. In an embodiment, the method oflowering body weight comprises administering a FGF-21 compound incombination with an exendin compound, wherein the FGF-21 compound isselected from the group consisting of human FGF-21, an FGF-21 analog andan FGF-21 derivative and wherein the exendin compound is selected fromthe group consisting of exendin-4, an exendin-4 analog, an exendin-4derivative, and an exendin-4 agonist. In a preferred embodiment, themethod of lowering body weight comprises administering a FGF-21 analogin combination with exendin-4. In another preferred embodiment, themethod of lowering body weight comprises administering a FGF-21 analogin combination with an exendin-4 analog. In another preferredembodiment, the method of lowering body weight comprises administering aFGF-21 analog and an exendin-4 derivative. In another preferredembodiment, the method of lowering body weight comprises administering aFGF-21 analog and an exendin-4 agonist.

The present invention also provides a method of treating obesitycomprising administering a FGF-21 compound in combination with a GLP-1compound. In more preferred embodiment, administering a FGF-21 compoundin combination with a GLP-1 compound results in a synergistic effect onweight loss. In an embodiment, the method of treating obesity comprisesadministering a FGF-21 compound in combination with a GLP-1 compound,wherein the FGF-21 compound is selected from the group consisting ofhuman FGF-21, an FGF-21 analog and an FGF-21 derivative and wherein theGLP-1 compound is selected from the group consisting of GLP-1 analog,GLP-1 derivative and GLP-1 fusion protein. In a preferred embodiment,the method of treating obesity comprises administering a FGF-21 analogin combination with a GLP-1 analog. In another preferred embodiment, themethod of treating obesity comprises administering a FGF-21 analog incombination with a GLP-1 derivative. In another preferred embodiment,the method of treating obesity comprises administering a FGF-21 analogand a GLP-1 fusion protein.

The present invention also provides a method of treating obesitycomprising administering a FGF-21 compound in combination with anexendin compound. In more preferred embodiment, administering a FGF-21compound in combination with an exendin compound results in asynergistic effect on weight loss. In an embodiment, the method oftreating obesity comprises administering a FGF-21 compound incombination with an exendin compound, wherein the FGF-21 compound isselected from the group consisting of human FGF-21, an FGF-21 analog andan FGF-21 derivative and wherein the exendin compound is selected fromthe group consisting of exendin-4, an exendin-4 analog, an exendin-4derivative, and exendin-4 agonist. In a preferred embodiment, the methodof treating obesity comprises administering a FGF-21 analog incombination with exendin-4. In another preferred embodiment, the methodof treating obesity comprises administering a FGF-21 analog incombination with an exendin-4 analog. In another preferred embodiment,the method of treating obesity comprises administering a FGF-21 analogand an exendin-4 derivative. In another preferred embodiment, the methodof treating obesity comprises administering a FGF-21 analog and anexendin-4 agonist.

The present invention also provides for the use of a FGF-21 compound anda GLP-1 compound in the manufacture of a medicament to lower bodyweight. The present invention also provides for the use of a FGF-21compound and a GLP-1 compound in the manufacture of a medicament tolower body weight, wherein the FGF-21 compound is selected from thegroup consisting of human FGF-21, an FGF-21 analog and an FGF-21derivative and wherein the GLP-1 compound is selected from the groupconsisting of a GLP-1 analog, a GLP-1 derivative and a GLP-1 fusionprotein. In a preferred embodiment, the present invention provides forthe use of a FGF-21 analog and a GLP-1 analog in the manufacture of amedicament to lower body weight. In another preferred embodiment, thepresent invention provides for the use of a FGF-21 analog and a GLP-1derivative in the manufacture of a medicament to lower body weight. Inanother preferred embodiment, the present invention provides for the useof a FGF-21 analog and a GLP-1 fusion protein in the manufacture of amedicament to lower body weight.

The present invention also provides for the use of a FGF-21 compound andan exendin compound in the manufacture of a medicament to lower bodyweight. The present invention also provides for the use of a FGF-21compound and an exendin compound in the manufacture of a medicament tolower body weight, wherein the FGF-21 compound is selected from thegroup consisting of human FGF-21, an FGF-21 analog and an FGF-21derivative and wherein the exendin compound is selected from the groupconsisting of exendin-4, an exendin-4 analog, an exendin-4 derivative,and an exendin-4 agonist. In a preferred embodiment, the presentinvention provides for the use of a FGF-21 analog and an exendin-4analog in the manufacture of a medicament to lower body weight. Inanother preferred embodiment, the present invention provides for the useof a FGF-21 analog and an exendin-4 derivative in the manufacture of amedicament to lower body weight. In another preferred embodiment, thepresent invention provides for the use of a FGF-21 analog and anexendin-4 agonist in the manufacture of a medicament to lower bodyweight.

The present invention also provides for the use of a FGF-21 compound anda GLP-1 compound in the manufacture of a medicament for the treatment ofobesity. The present invention also provides for the use of a FGF-21compound and a GLP-1 compound in the manufacture of a medicament for thetreatment of obesity, wherein the FGF-21 compound is selected from thegroup consisting of human FGF-21, an FGF-21 analog and an FGF-21derivative and wherein the GLP-L compound is selected from the groupconsisting of a GLP-1 analog, a GLP-1 derivative and a GLP-1 fusionprotein. In a preferred embodiment, the present invention provides forthe use of a FGF-21 analog and a GLP-1 analog in the manufacture of amedicament for the treatment of obesity. In another preferredembodiment, the present invention provides for the use of a FGF-21analog and a GLP-1 derivative in the manufacture of a medicament for thetreatment of obesity. In another preferred embodiment, the presentinvention provides for the use of a FGF-21 analog and a GLP-1 fusionprotein in the manufacture of a medicament for the treatment of obesity.

The present invention also provides for the use of a FGF-21 compound andan exendin compound in the manufacture of a medicament to treat obesity.The present invention also provides for the use of a FGF-21 compound andan exendin compound in the manufacture of a medicament to lower bodyweight wherein the FGF-21 compound is selected from the group consistingof human FGF-21, an FGF-21 analog and an FGF-21 derivative and whereinthe exendin compound is selected from the group consisting of exendin-4,an exendin-4 analog, an exendin-4 derivative, and an exendin-4 agonist.In a preferred embodiment, the present invention provides for the use ofa FGF-21 analog and an exendin-4 analog in the manufacture of amedicament to treat obesity. In another preferred embodiment, thepresent invention provides for the use of a FGF-21 analog and anexendin-4 derivative in the manufacture of a medicament to treatobesity. In another preferred embodiment, the present invention providesfor the use of a FGF-21 analog and an exendin-4 agonist in themanufacture of a medicament to treat obesity.

The FGF-21 compounds of the present invention may be human FGF-21, aFGF-21 analog or a FGF-21 derivative. In a preferred embodiment, theFGF-21 compound of the present invention is a FGF-21 analog in a morepreferred embodiment, the present invention provides FGF-21 compoundscomprising one or two engineered disulfide bonds. In a more preferredembodiment, the present invention provides FGF-21 compounds whichcomprise an amino acid sequence comprising a cysteine substitution atpositions 21, 26, 33, 118, 119, 121, 122, or 134 of FGF-21 (SEQ ID NO:2). In another preferred embodiment, the FGF-21 compound comprises anamino acid sequence comprising an amino acid substitution at position167 of FGF-21 (SEQ ID NO: 2), wherein the substitution is not Ser orTyr. In another preferred embodiment, the FGF-2 compound comprises anamino acid sequence comprising an amino acid substitution at position121 of FGF-21 (SEQ ID NO: 2), wherein the substitution is any amino acidexcept Gln or Asn. In a more preferred embodiment, the amino acid atposition 121 of FGF-21 (SEQ ID NO: 2) is selected from the groupconsisting of Ala, Val, Ser, Asp, or Glu. In a more preferredembodiment, the FGF-21 compound comprises an amino acid sequenceselected from the group consisting of Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO:9), Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10),Cys²¹-Cys³³-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 11), Cys²⁶-Cys¹²²-Ala¹⁶⁷-FGF-21(SEQ ID NO: 12), and Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO:13).In a more preferred embodiment, the FGF-21 compound consists of an aminoacid sequence selected from the group consisting of Cys¹¹⁸-Cys¹³⁴-FGF-21(SEQ ID NO: 9), Cys²⁶-Cys¹²²-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10),Cys²¹-Cys³³-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 11), Cys²⁶-Cys¹²²-Ala¹⁶⁷-FGF-21(SEQ ID NO: 12), and Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO:13).

In an embodiment, the present invention provides FGF-21 compounds whichconsist of a cysteine substitution at positions 21, 26, 33, 118, 119,121, 122, or 134 of FGF-21 (SEQ ID NO: 2). In another embodiment, theFGF-21 compound consist of an amino acid substitution at position 167 ofFGF-21 (SEQ ID NO: 2), wherein the substitution is not Ser or Tyr. Inanother preferred embodiment, the FGF-21 compound consists of asubstitution at position 121 of FGF-21 (SEQ ID NO: 2), wherein thesubstitution is any amino acid except Gln or Asn. In a more preferredembodiment, the amino acid at position 121 of FGF-21 (SEQ ID NO: 2) isselected from the group consisting of Ala, Val, Ser, Asp, or Glu. In amore preferred embodiment, the FGF-21 compound consists of an amino acidsequence selected from the group consisting of Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQID NO: 9), Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10),Cys²¹-Cys³³-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 11), Cys²⁶-Cys¹²²-Ala¹⁶⁷-FGF-21(SEQ ID NO: 12), and Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO; 13).

Further, the present invention provides GLP-1 compounds to be used incombination with the FGF-21 compounds of the present invention. In anembodiment, the GLP-1 compound is a GLP-1 analog, GLP-1 derivative, or aGLP-1 fusion protein. In a more preferred embodiment, the GLP-1 compoundis a GLP-1 analog or derivative. In a preferred embodiment, the GLP-1compound is a GLP-1 analog. In a more preferred embodiment, the GLP-1compound comprises of an amino acid sequence of SEQ ID NO:

His Xaa Glu Gly Thr Phe Thr Ser Asp Val Ser Ser            10                  15Tyr Leu Glu Xaa Gln Ala Ala Lys Glu Phe Ile Ala    20                  25                  30Trp Leu Xaa Xaa Gly Xaa Xaa Xaa Xaa Xaa Xaa Xaa                35                  40 Xaa Xaa Xaa         45whereinXaa at position 8 is Gly, Ala, Val,Xaa at position 22 is Gly, Glu, Asp, or Lys,Xaa at position 33 is Val, or Ile,Xaa at position 34 is Lys or Arg,Xaa at position 36 is Arg or Gly,Xaa at position 37 is selected from the group consisting of NH₂, Gly andPro,Xaa at position 38 is Ser or absent,Xaa at position 39 is Ser or absent,Xaa at position 40 is Gly or absent,Xaa at position 41 is Ala or absent,Xaa at position 42 is Pro or absent,Xaa at position 43 is Pro or absent,Xaa at position 44 is Pro or absent, andXaa at position 45 is Ser or absent.

In an embodiment, the GLP-1 compound comprises Ala at Xaa₈. In anotherembodiment, the GLP-1 compound comprises Val at Xaa₈. In anotherembodiment, the GLP-1 compound comprises Gly at Xaa₂₂. In a preferredembodiment, the GLP-1 compound comprises Gly at Xaa₂₂. In a preferredembodiment, the GLP-1 compound comprises Lys at Xaa₃₄. In anotherpreferred embodiment, the GLP-1 compound comprises Arg at Xaa₃₄. In amore preferred embodiment, the amino acid sequence of the GLP-1 analogcomprisesVal⁸-Glu²²-Ile³³-Gly³⁶-Pro³⁷-Ser³⁸-Ser³⁹-Gly⁴⁰-Ala⁴¹-Pro⁴²-Pro⁴³-Pro⁴⁴-Ser⁴⁵-GLP-1(SEQ ID NO: 19). In another embodiment, the amino acid sequence of theGLP-1 analog consists ofVal⁸-Glu²²-Ile³³-Gly³⁶-Pro³⁷-Ser³⁸-Ser³⁹-Gly⁴⁰-Ala⁴¹-Pro⁴²-Pro⁴³-Pro⁴⁴-Ser⁴⁵-GLP-1(SEQ ID NO: 19).

In a preferred embodiment, the amino acid sequence of the GLP-1 compoundcomprises Val⁸-GLP-1 (SEQ ID NO: 15). In a preferred embodiment, theamino acid sequence of the GLP-1 compound consists of the amino acidsequence Val⁸-GLP-1 (SEQ ID NO: 15). In another preferred embodiment,the amino acid sequence of the GLP-1 compound comprises Arg³⁴-GLP-1 (SEQID NO: 16). In another preferred embodiment, the amino acid sequence ofthe GLP-1 compound consists of Arg³⁴-GLP-1 (SEQ ID NO: 16). In anotherpreferred embodiment, the amino acid sequence of the GLP-1 compoundcomprises Val⁸-Glu²²-GLP-1 (SEQ ID NO: 17). In another preferredembodiment, the amino acid sequence of the GLP-1 compound consists ofVal⁸-Glu²²-GLP-1 (SEQ ID NO: 17). In another preferred embodiment, theamino acid sequence of Val⁸-Glu²²-GLP-1 (SEQ ID NO: 17) comprises offive additional amino acid sequence substitutions. In another preferredembodiment, the amino acid sequence of Val⁸-Glu²²-GLP-1 (SEQ ID NO: 17)consists of five additional amino acid sequence substitutions. Inanother preferred embodiment, the amino acid sequence ofVal⁸-Glu²²-GLP-1 (SEQ ID NO: 17) comprises of four additional amino acidsequence substitutions. In another preferred embodiment, the amino acidsequence of Val⁸-Glu²²-GLP-1 (SEQ ID NO: 17) consists of four additionalamino acid sequence substitutions. In another preferred embodiment, theamino acid sequence of Val⁸-Glu²²-GLP-1 (SEQ ID NO: 17) comprises ofthree additional amino acid sequence substitutions. In another preferredembodiment, the amino acid sequence of Val⁸-Glu²²-GLP-1 (SEQ ID NO: 17)consists of three additional amino acid sequence substitutions. Inanother preferred embodiment, the amino acid sequence ofVal⁸-Glu²²-GLP-1 (SEQ ID NO: 17) comprises of two additional amino acidsequence substitutions. In another preferred embodiment, the amino acidsequence of Val⁸-Glu²²-GLP-1 (SEQ ID NO: 17) consists of two additionalamino acid sequence substitutions. In another preferred embodiment, theamino acid sequence of Val⁸-Glu²²-GLP-1 (SEQ ID NO: 17) comprises of oneadditional amino acid sequence substitution In another preferredembodiment, the amino acid sequence of Val⁸-Glu²²-GLP-1 (SEQ ID NO: 17)consists of one additional amino acid sequence substitution.

The present invention also provides GLP-1 derivatives in combinationwith FGF-21 compounds. In another preferred embodiment, the GLP-1derivative is pegylated. In a preferred embodiment, the GLP-1 derivativecomprises of an amino acid sequence of SEQ ID NO: 20

His Xaa Glu Gly Thr Phe Thr Ser Asp Val Ser Ser           10                  15Tyr Leu Glu Xaa Gln Ala Ala Lys Glu Phe Ile Ala    20                  25                  30Trp Leu Xaa Lys Gly Gly Pro Ser Ser Gly Ala Pro                35                  40 Pro Pro Cys Xaa         45whereinXaa at position 8 is: D-Ala, Gly, Val, Leu, Ile, Ser, or Thr;Xaa at position 22 is: Gly, Glu, Asp, or Lys;Xaa at position 33 is: Val or IleXaa at position 46 is: Cys or Cys-NH₂and wherein one PEG molecule is covalently attached to Cys⁴⁵ and one PEGmolecule is covalently attached to Cys⁴⁶ or Cys⁴⁶-NH₂. In anotherembodiment, the GLP-1 derivative consists of the amino acid sequence ofSEQ ID NO: 20.

In an embodiment, Xaa₈ is Val or Gly. In another embodiment, Xaa₂₂ isGly or Glu. In another embodiment, Xaa₃₃ is Ile. In an embodiment, Xaa₄is Cys-NH₂. In a preferred embodiment, the GLP-1 derivative comprises ofthe amino acid sequence of Val⁸-Glu²²-Ile³³-Cys-NH₂ ⁴⁶-GLP-1 (SEQ ID NO:21). In a preferred embodiment, the GLP-1 derivative consists of theamino acid sequence of Val⁸-Glu²²-Ile³³-Cys-NH₂ ⁴⁶-GLP-1 (SEQ ID NO:21).

In another embodiment, the GLP-1 compound is a GLP-1 fusion protein. Ina preferred embodiment, the GLP-1 fusion protein comprises a GLP-1portion and an Fc portion of an immunoglobulin. In a preferredembodiment, the GLP-t fusion protein comprises a GLP-1 analog and the Fcportion of an immunoglobulin wherein the GLP-1 analog comprises an aminoacid sequence of SEQ ID NO: 22

His Xaa Glu Gly Thr Phe Thr Ser Asp Val Ser Ser            10                  15 Tyr Leu Glu Glu Gln Ala Ala Lys Glu Phe Ile Ala    20                  25                  30Trp Leu Xaa Xaa Gly Gly Xaa                 35whereinXaa at position 8 is Gly or Val;Xaa at position 33 is Val or Lys;Xaa at position 34 is Lys or Asn;Xaa at position 37 is Gly, Pro or is absent,and wherein the GLP analog is fused to the Fc portion of animmunoglobulin comprising the amino acid sequence of Formula IV (SEQ IDNO: 23)

Ala Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys                5                   10Pro Ala Pro Xaa Xaa Xaa Gly Gly Pro Ser Val Phe        15                  20 Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile25                  30                  35Ser Arg Thr Pro Gln Val Thr Cys Val Val Val Asp            40                  45 Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp    50                  55                  60Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr                65                  70 Lys Pro Arg Glu Glu Gln Phe Xaa Ser Thr Tyr Arg        75                  80 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp85                  90                  95Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn            100                 105Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser    110                 115                 120Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr               125                  130 Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn        135                 140Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr145                 150                 155Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly            160                 165Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val    170                 175                 180Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg                185                 190Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn        195                 200Val Phe Ser Cys Ser Val Met His Glu Ala Leu His205                 210                 215Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu            220                 225 Gly Xaa     230wherein:Xaa at position 16 is Pro or Glu;Xaa at position 17 is Phe, Val, or Ala;Xaa at position 18 is Leu, Glu, or Ala;Xaa at position 80 is Asn or Ala; andXaa at position 230 is Lys or is absent.

In a preferred embodiment, the C-terminus of the GLP-1 analog and theN-terminus of the Fc portion of an immunoglobulin are preferably fusedtogether via 1, 1.5 (SEQ ID NO: 7) or 2 repeats (SEQ ID NO: 8) of aG-rich peptide linker having the sequenceGly-Gly-Gly-Gly-Ser-Gly-Gly-Gly-Gly-Ser-Gly-Gly-Gly-Gly-Ser (SEQ ID NO:6).

In a preferred embodiment, the GLP-1 fusion protein comprises a GLP-1analog and the Fc portion of an immunoglobulin wherein the GLP-1 analogconsists of an amino acid sequence of SEQ ID NO: 22 and wherein the GLPanalog is fused to the Fc portion of an immunoglobulin consisting of theamino acid sequence of SEQ ID NO: 23. In another embodiment, the GLP-1fusion protein further comprises a linker. In another embodiment, theGLP-1 fusion protein further comprises a linker, wherein the linkercomprises an amino acid sequence is selected from the group consistingof SEQ ID NO: 6, SEQ ID NO:7 and SEQ ID NO:8. In another embodiment, theGLP-1 fusion protein further comprises a linker, wherein the linkerconsists of an amino acid sequence is selected from the group consistingof SEQ ID NO: 6, SEQ ID NO:7 and SEQ ID NO:8. In an embodiment, thelinker comprises an amino acid sequence of SEQ ID NO:6. In anembodiment, the linker comprises an amino acid sequence of SEQ ID NO:7.In an embodiment, the linker comprises an amino acid sequence of SEQ IDNO: 8. In an embodiment, the linker consists of an amino acid sequenceof SEQ ID NO:6. In an embodiment, the linker consists of an amino acidsequence of SEQ ID NO:7. In an embodiment, the linker consists of anamino acid sequence of SEQ ID NO:8

In an embodiment of the present invention, the GLP-1 portion comprisesGly at Xaa₈. In another embodiment, the GLP-1 portion comprises Val atXaa₈. In an embodiment, the GLP-1 portion comprises Gly at Xaa₂₂. In apreferred embodiment, the GLP-1 portion comprises Glu at Xaan. In anembodiment, the GLP-1 portion comprises Lys at Xaa₃₄. In anotherembodiment, the GLP-1 portion comprises Asn at Xaa₃₄. In an embodiment,Xaa₃₇ of the GLP-1 portion is absent. In a preferred embodiment, theGLP-1 portion comprises Gly at Xaa₃₇. In another preferred embodiment,the GLP-1 portion comprises Pro at Xaa₃₇.

Preferred GLP-1 fusion proteins of the present invention include thefollowing proteins: Gly⁸-Glu²²-Gly³⁶-GLP-1-1L-IgG4 (S228P) (SEQ ID NO:24), Gly⁸-Glu²²-Gly³⁶-GLP-1-1L-IgG4 (S228P, F234A, L235A) (SEQ ID NO:25), Gly⁸-Glu²²-Gly³⁶-GLP-1-1L-IgG4 (S228P, N297A) (SEQ ID NO: 26),Gly⁸-Glu²²-Gly³⁶-GLP-1-IgG4 (S228P, F234A, L235A, N297A) (SEQ ID NO:27), Gly⁸-Glu²²-Gly³⁶-GLP-1L-IgG4 (S228P, des K) (SEQ ID NO: 28),Gly⁸-Glu²²-Gly³⁶-GLP-1-1L-IgG4 (S228P, F234A, L235A, des K) (SEQ ID NO:29), Gly⁸-Glu²²-Gly³⁶-GLP-1-1 L-IgG4 (S228P, N297A,des K) (SEQ ID NO:30), Gly⁸-Glu²²-Gly³⁶-GLP-1-1L-IgG4 (S228P, F234A, L235A, N297A, des K)(SEQ ID NO: 31), Gly⁸-Glu²²-Gly³⁶-GLP-1-1.5L-LgG4 (S228P) (SEQ ID NO:32), Gly⁸-Glu²²-Gly³⁶-GLP-1-1.5L-IgG4 (S228P, F234A, L235A) (SEQ ID NO:33), Gly⁸-Glu²²-Gly³⁶-GLP-1-1.5L-IgG4 (S228P, N297A) (SEQ ID NO: 34),Gly⁸-Glu²²-Gly³⁶-GLP-1-1.5L-IgG4 (S228P, F234A, L235A, N297A) (SEQ IDNO: 35), Gly⁸-Glu²²-Gly³⁶-GLP-1-1.5L-IgG4 (S228P, des K) (SEQ ID NO:36), Gly⁸-Glu²²-Gly³⁶-GLP-1-1.5L-IgG4 (S228P, F234A, L235A, des K) (SEQID NO: 37), Gly⁸-Glu²²-Gly³⁶-GLP-1-1.5L-IgG4 (S228P, N297A des K) (SEQID NO: 38), Gly⁸-Glu²²-Gly³⁶-GLP-1-1.5L-IgG4 (S228P, F234A, L235A,N297A, des K) (SEQ ID NO: 39), Gly⁸-Glu²²-Gly³⁶-GLP-1-2L-IgG4 (S228P)(SEQ ID NO: 40), Gly⁸-Glu²²-Gly³⁶-GLP-1-2L-IgG4 (S228P, F234A, L235A)(SEQ ID NO: 41), Gly⁸-Glu²²-Gly³⁶-GLP-1-2L-IgG4 (S228P, N297A) (SEQ IDNO: 42), Gly⁸-Glu²²-Gly³⁶-GLP-1-2L-IgG4 (S228P, F234A, L235A, N297A)(SEQ ID NO: 43), (Gly⁸-Glu²²-Gly³⁶-GLP-1-2L-IgG4 (S228P, des K) (SEQ IDNO: 44), Gly⁸-Glu²²-Gly³⁶-GLP-1-2L-IgG4 (S228P, F234A, L235A, des K)(SEQ ID NO: 45), Gly⁸-Glu²²-Gly³⁶-GLP-1-2L-IgG4 (S228P, N297A, des K)(SEQ ID NO: 46), Gly⁸-Glu²²-Gly³⁶-GLP-1-2L-IgG4 (S228P, F234A, L235A,N297A, des K) (SEQ ID NO: 47), and the Val⁸ forms of all of the above.

The present invention also includes FGF-21 compounds in combination withGLP-1 compounds, wherein the GLP-1 compound comprises an amino acidsequence selected from the group consisting of SEQ ID NO: 47, SEQ ID NO:48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ IDNO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, and SEQ ID NO: 57.The present invention also includes FGF-21 compounds in combination withGLP-1 compounds, wherein the GLP-1 compound consists of an amino acidsequence selected from the group consisting of SEQ ID NO: 47, SEQ ID NO:48, SEQ ID NO: 49, SEQ ID NO: 50. SEQ ID NO: 51, SEQ ID NO: 52, SEQ IDNO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, and SEQ ID NO: 57.

Preferred FGF-21 compound and GLP-1 compound combinations of the presentinvention include. Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combinationwith Val⁸-GLP-1 (SEQ ID NO: 15), Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) incombination with Val⁸-Glu²²-GLP-1 (SEQ ID NO: 17), Cys¹¹⁸-Cys¹³⁴-FGF-21(SEQ ID NO: 9) in combination with isVal⁸-Glu²²-Ile³³-Gly³⁶-Pro³⁷-Ser³⁸-Ser³⁹-Gly⁴⁰-Ala⁴¹-Pro⁴²-Pro⁴³-Pro⁴⁴-Ser⁴⁵-GLP-1(SEQ ID NO: 19), Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination withpegylated Val⁸-Glu²²-Ile³³-Cys-NH₂-GLP-1 (SEQ ID NO: 21),Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination with Arg-GLP-1 (SEQID NO: 16), Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination withacylated Arg³⁴-GLP-1 (SEQ ID NO: 16), Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO:9) in combination with Arg³⁴-Lys²⁶-(N-ε-(γ-Glu(N-α-hexadecanoyl)))-GLP-1 (SEQ ID NO: IS), Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ IDNO: 9) in combination with Gly⁸-Glu²²-Gly³⁶-GLP-1-1L-IgG4 (S228P) (SEQID NO: 24), Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1-1L-IgG4 (S228P, F234A, L235A) (SEQ ID NO: 25),Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1-L-IgG4 (S228P, N297A) (SEQ ID NO: 26),Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1-L-IgG4 (S228P, F234A, L235A, N297A) (SEQ ID NO:27), Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1-1 L-IgG4 (S228P, des K) (SEQ ID NO: 28),Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1-1L-IgG4 (S228P, F234A, L235A, des K) (SEQ ID NO:29), Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1-1L-IgG4 (S228P, N297A, des K) (SEQ ID NO: 30).Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1-1L-IgG4 (S228P, F234A, L235A, N297A, des K) (SEQID NO: 31), Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1-1.5L-IgG4 (S228P) (SEQ ID NO: 32),Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1-1.5L-IgG4 (S228P, F234A, L235A) (SEQ ID NO: 33),Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1-1.5L-IgG4 (S228P, N297A) (SEQ ID NO: 34),Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1-1.5L-IgG4 (S228P, F234A, L235A, N297A) (SEQ IDNO: 35), Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1-1.5L-IgG4 (S228P, des K) (SEQ ID NO: 36),Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1-1.5L-IgG4 (S228P, F234A, L235A, des K) (SEQ IDNO: 37), Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1-1.5L-IgG4 (S228P, N297A, des K) (SEQ ID NO: 38),Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1-1.5L-IgG4 (S228P, F234A, L235A, N297A, des K)(SEQ ID NO: 39), Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1-2L-IgG4 (S228P) (SEQ ID NO: 40),Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1-2L-IgG4 (S228P, F234A, L235A) (SEQ ID NO: 41),Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1-2L-IgG4 (S228P, N297A) (SEQ ID NO: 42),Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1-2L-IgG4 (S228P, F234A, L235A, N297A) (SEQ ID NO:43), Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1-2L-IgG4 (S228P, des K) (SEQ ID NO: 44),Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1-2L-IgG4 (S228P, F234A, L235A, des K) (SEQ ID NO:45), Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1-2L-IgG4 (S228P, N297A, des K) (SEQ ID NO: 46),Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1-2L-IgG4 (S228P, F234A, L235A, N297A, des K) (SEQID NO: 47), Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination withexendin-4 (SEQ ID NO: 4), Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) incombination with albiglutide (SEQ ID NO: 48). Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQID NO: 9) in combination with a GLP-1 compound comprising the amino acidsequence of SEQ ID NO: 47, Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) incombination with a GLP-1 compound consisting of the amino acid sequenceof SEQ ID NO: 47, Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combinationwith a GLP-1 compound comprising the amino acid sequence of SEQ ID NO:48, Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination with a GLP-1compound consisting of the amino acid sequence of SEQ ID NO: 48,Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination with a GLP-1 compoundcomprising the amino acid sequence of SEQ ID NO: 49,Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination with a GLP-1 compoundconsisting of the amino acid sequence of SEQ ID NO: 49,Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination with a GLP-1 compoundcomprising the amino acid sequence of SEQ ID NO: 50,Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination with a GLP-1 compoundconsisting of the amino acid sequence of SEQ ID NO: 50,Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination with a GLP-1 compoundcomprising the amino acid sequence of SEQ ID NO: 51,Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination with a GLP-1 compoundconsisting of the amino acid sequence of SEQ ID NO: 51,Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination with a GLP-1 compoundcomprising the amino acid sequence of SEQ ID NO: 52,Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination with a GLP-1 compoundconsisting of the amino acid sequence of SEQ ID NO: 52,Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination with a GLP-1 compoundcomprising the amino acid sequence of SEQ ID NO: 53,Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination with a GLP-1 compoundconsisting of the amino acid sequence of SEQ ID NO: 53,Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination with a GLP-1 compoundcomprising the amino acid sequence of SEQ ID NO: 54,Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination with a GLP-1 compoundconsisting of the amino acid sequence of SEQ ID NO: 54,Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination with a GLP-1 compoundcomprising the amino acid sequence of SEQ ID NO: 55,Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination with a GLP-1 compoundconsisting of the amino acid sequence of SEQ ID NO: 55,Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination with a GLP-1 compoundcomprising the amino acid sequence of SEQ ID NO: 56,Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination with a GLP-1 compoundconsisting of the amino acid sequence of SEQ ID NO: 56,Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination with a GLP-1 compoundcomprising the amino acid sequence of SEQ ID NO: 57, andCys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination with a GLP-1 compoundconsisting of the amino acid sequence of SEQ ID NO: 57

In a preferred embodiment, the combination of the present inventioncomprises an FGF-21 analog comprising the amino acid sequence ofCys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination with a GLP-1 analog,wherein the GLP-1 analog comprises the amino acid sequence ofVal⁸-Glu²²-GLP-1 (SEQ ID NO: 17) with five additional amino acidsequence substitutions. In a preferred embodiment, the combination ofthe present invention comprises an FGF-21 analog consisting of the aminoacid sequence of Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination witha GLP-1 analog, wherein the GLP-1 analog consists of the amino acidsequence of Val⁸-Glu²²-GLP-1 (SEQ ID NO: 17) with five additional aminoacid sequence substitutions. In another preferred embodiment, thecombination of the present invention comprises an FGF-21 analogcomprising the amino acid sequence of Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO:9) in combination with a GLP-1 analog, wherein the GLP-1 analogcomprises the amino acid sequence of Val⁸-Glu²²-GLP-1 (SEQ ID NO: 17)with four additional amino acid sequence substitutions. In anotherpreferred embodiment, the combination of the present invention comprisesan FGF-21 analog consisting of the amino acid sequence ofCys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination with a GLP-1 analog,wherein the GLP-1 analog consists of the amino acid sequence ofVal⁸-Glu²²-GLP-1 (SEQ ID NO: 17) with four additional amino acidsequence substitutions. In another preferred embodiment, the combinationof the present invention comprises an FGF-2 analog comprising the aminoacid sequence of Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination witha GLP-1 analog, wherein the GLP-1 analog comprises the amino acidsequence of Val⁸-Glu²²-GLP-1 (SEQ ID NO: 17) with three additional aminoacid sequence substitutions. In another preferred embodiment, thecombination of the present invention comprises an FGF-21 analogconsisting of the amino acid sequence of Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ IDNO: 9) in combination with a GLP-1 analog, wherein the GLP-1 analogconsists of the amino acid sequence of Val⁸-Glu²²-GLP-1 (SEQ ID NO: 17)with three additional amino acid sequence substitutions. In anotherpreferred embodiment, the combination of the present invention comprisesan FGF-21 analog comprising the amino acid sequence ofCys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination with a GLP-1 analog,wherein the GLP-1 analog comprises the amino acid sequence ofVal⁸-Glu²²-GLP-1 (SEQ ID NO: 17) with two additional amino acid sequencesubstitutions. In another preferred embodiment, the combination of thepresent invention comprises an FGF-21 analog consisting of the aminoacid sequence of Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination witha GLP-1 analog, wherein the GLP-1 analog consists of the amino acidsequence of Val⁸-Glu²²-GLP-1 (SEQ ID NO: 17) with two additional aminoacid sequence substitutions. In another preferred embodiment, thecombination of the present invention comprises an FGF-21 analogcomprising the amino acid sequence of Cys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO:9) in combination with a GLP-1 analog, wherein the GLP-1 analogcomprises the amino acid sequence of Val⁸-Glu²²-GLP-1 (SEQ ID NO: 17)with one additional amino acid sequence substitution. In anotherpreferred embodiment, the combination of the present invention comprisesan FGF-21 analog consisting of the amino acid sequence ofCys¹¹⁸-Cys¹³⁴-FGF-21 (SEQ ID NO: 9) in combination with a GLP-1 analog,wherein the GLP-1 analog consists of the amino acid sequence ofVal⁸-Glu²²-GLP-1 (SEQ ID NO: 17) with one additional amino acid sequencesubstitution.

More preferred FGF-21 compound and GLP-1 compound combinations of thepresent invention include Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) incombination with Val⁸-GLP-1 (SEQ ID NO: 17), Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21(SEQ ID NO: 10) in combination with Val⁸-Glu²²-GLP-1 (SEQ ID NO: 17),Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) in combination with isVal⁸-Glu²²-Ile³³-Gly³⁶-Pro³⁷-Ser³⁸-Ser³⁹-Gly⁴⁰-Ala⁴¹-Pro⁴²-Pro⁴³-Pro⁴⁴-Ser⁴⁵-GLP-1(SEQ ID NO: 19), Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) incombination with pegylated Val⁸-Glu²²-Ile³³-Cys-NH₂-GLP-1 (SEQ ID NO:21), Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) in combination withArg³⁴-GLP-1 (SEQ ID NO: 16), Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10)in combination with acylated Arg-GLP-1 (SEQ ID NO: 16),Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) in combination withArg³⁴-Lys²⁶-(N-ε-(γ-Glu (N-α-hexadecanoyl)))-GLP-1 (SEQ ID NO: 18),Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1(7-37)-1L-IgG4 (S228P) (SEQ ID NO: 24),Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1(7-37)-1L-IgG4 (S228P, F234A, L235A) (SEQ ID NO:25), Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1(7-37)-1L-IgG4 (S228P, N297A) (SEQ ID NO: 26),Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1(7-37)-1L-IgG4 (S228P, F234A, L235A, N297A) (SEQID NO: 27), Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) in combinationwith Gly⁸-Glu²²-Gly³⁶-GLP-1(7-37)-1L-IgG4 (S228P, des K) (SEQ ID NO:28), Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1(7-37)-1L-IgG4 (S228P, F234A, L235A, des K) (SEQID NO: 29), Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) in combinationwith Gly⁸-Glu²²-Gly³⁶-GLP-1(7-37)-1L-IgG4 (S228P, N297A, des K) (SEQ IDNO: 30), Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO:10) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1(7-37)-1L-IgG4 (S228P, F234A, L235A, N297A, des K)(SEQ ID NO: 31), Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) incombination with Gly⁸-Glu²²-Gly³⁶-GLP-1(7-37)-1.5L-IgG4 (S228P) (SEQ IDNO: 32), Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO:10) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1(7-37)-1.5L-IgG4 (S228P, F234A, L235A) (SEQ ID NO:33), Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1(7-37)-1.5L-IgG4 (S228P, N297A) (SEQ ID NO: 34),Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO:10) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1-1.5L-IgG4 (S228P, F234A, L235A, N297A) (SEQ IDNO: 35), Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1(7-37)-1.5L-IgG4 (S228P, des K) (SEQ ID NO: 36),Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1(7-37)-1.5L-IgG4 (S228P, F234A, L235A, des K) (SEQID NO: 37), Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) in combinationwith Gly⁸-Glu²²-Gly³⁶-GLP-1(7-37)-1.5L-IgG4 (S228P, N297A, des K) (SEQID NO: 38), Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) in combinationwith Gly⁸-Glu²²-Gly³⁶-GLP-1-1.5L-IgG4 (S228P, F234A, L235A, N297A, desK) (SEQ ID NO: 39), Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) incombination with Gly⁸-Glu²²-Gly³⁶-GLP-1-2L-IgG4 (S228P) (SEQ ID NO: 40),Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO:10) in combination with(Gly⁸-Glu²²-Gly³⁶-GLP-1-2L-IgG4 (S228P, F234A, L235A) (SEQ ID NO: 41),Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1-2L-IgG4 (S228P, N297A) (SEQ ID NO: 42),Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO:10) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1 (7-37)-2L-IgG4 (S228P, F234A, L235A, N297A) (SEQID NO: 43), Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO:10) in combinationwith Gly⁸-Glu²²-Gly³⁶-GLP-1-2L-IgG4 (S228P, des K) (SEQ ID NO 44),Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1-2L-IgG4 (S228P, F234A, L235A, des K) (SEQ ID NO:45), Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1-2L-IgG4 (S228P, N297A, des K) (SEQ ID NO: 46),Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO:10) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1(7-37)-2L-IgG4 (S228P, F234A, L235A, N297A, des K)(SEQ ID NO: 47), Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO:10) incombination with exendin-4 (SEQ ID NO: 4), Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21(SEQ ID NO: 10) in combination with albiglutide (SEQ ID NO: 48).Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) in combination with a GLP-1compound comprising the amino acid sequence of SEQ ID NO: 47,Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO:10) in combination with a GLP-1compound consisting of the amino acid sequence of SEQ ID NO: 47,Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) in combination with a GLP-1compound comprising the amino acid sequence of SEQ ID NO: 48,Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) in combination with a GLP-1compound consisting of the amino acid sequence of SEQ ID NO: 48,Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) in combination with a GLP-1compound comprising the amino acid sequence of SEQ ID NO: 49,Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO:10) in combination with a GLP-1compound consisting of the amino acid sequence of SEQ ID NO: 49.Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) in combination with a GLP-1compound comprising the amino acid sequence of SEQ ID NO: 50,Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) in combination with a GLP-1compound consisting of the amino acid sequence of SEQ ID NO: 50,Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO:10) in combination with a GLP-1compound comprising the amino acid sequence of SEQ ID NO: 51,Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) in combination with a GLP-1compound consisting of the amino acid sequence of SEQ ID NO: 51,Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) in combination with a GLP-1compound comprising the amino acid sequence of SEQ ID NO: 52,Cys¹¹⁸-Cys¹¹⁸-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) in combination with a GLP-1compound consisting of the amino acid sequence of SEQ ID NO: 52,Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) in combination with a GLP-1compound comprising the amino acid sequence of SEQ ID NO: 53,Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) in combination with a GLP-1compound consisting of the amino acid sequence of SEQ ID NO: 53,Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) in combination with a GLP-1compound comprising the amino acid sequence of SEQ ID NO: 54,Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) in combination with a GLP-1compound consisting of the amino acid sequence of SEQ ID NO: 54,Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO:10) in combination with a GLP-1compound comprising the amino acid sequence of SEQ ID NO: 55,Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) in combination with a GLP-1compound consisting of the amino acid sequence of SEQ ID NO: 55,Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO:10) in combination with a GLP-1compound comprising the amino acid sequence of SEQ ID NO: 56,Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) in combination with a GLP-1compound consisting of the amino acid sequence of SEQ ID NO: 56,Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO:10) in combination with a GLP-1compound comprising the amino acid sequence of SEQ ID NO: 57 andCys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO:10) in combination with a GLP-1compound consisting of the amino acid sequence of SEQ ID NO: 57.

In a preferred embodiment, the combination of the present inventioncomprises an FGF-21 analog comprising the amino acid sequence ofCys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) in combination with a GLP-1analog, wherein the GLP-1 analog comprises the amino acid sequence ofVal⁸-Glu²²-GLP-1 (SEQ ID NO: 17) with five additional amino acidsequence substitutions. In a preferred embodiment, the combination ofthe present invention comprises an FGF-21 analog consisting of the aminoacid sequence of Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) incombination with a GLP-1 analog, wherein the GLP-1 analog consists ofthe amino acid sequence of Val⁸-Glu²²-GLP-1 (SEQ ID NO: 17) with fiveadditional amino acid sequence substitutions. In another preferredembodiment, the combination of the present invention comprises an FGF-21analog comprising the amino acid sequence of Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21(SEQ ID NO: 10) in combination with a GLP-1 analog, wherein the GLP-1analog comprises the amino acid sequence of Val⁸-Glu²²-GLP-1 (SEQ ID NO:17) with four additional amino acid sequence substitutions. In anotherpreferred embodiment, the combination of the present invention comprisesan FGF-21 analog consisting of the amino acid sequence ofCys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) in combination with a GLP-1analog, wherein the GLP-1 analog consists of the amino acid sequence ofVal⁸-Glu²²-GLP-1 (SEQ ID NO: 17) with four additional amino acidsequence substitutions. In another preferred embodiment, the combinationof the present invention comprises an FGF-21 analog comprising the aminoacid sequence of Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) incombination with a GLP-1 analog, wherein the GLP-1 analog comprises anamino acid sequence of Val⁸-Glu²²-GLP-1 (SEQ ID NO: 17) with threeadditional amino acid sequence substitutions. In another preferredembodiment, the combination of the present invention comprises an FGF-21analog consisting of the amino acid sequence ofCys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) in combination with a GLP-1analog, wherein the GLP-1 analog consist of an amino acid sequence ofVal⁸-Glu²²-GLP-1 (SEQ ID NO: 17) with three additional amino acidsequence substitutions. In another preferred embodiment, the combinationof the present invention comprises an FGF-21 analog comprising the aminoacid sequence of Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) incombination with a GLP-1 analog, wherein the GLP-1 analog comprises theamino acid sequence of Val⁸-Glu²²-GLP-1 (SEQ ID NO: 17) with twoadditional amino acid sequence substitutions. In another preferredembodiment, the combination of the present invention comprises an FGF-21analog consisting of the amino acid sequence ofCys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) in combination with a GLP-1analog, wherein the GLP-1 analog consists of the amino acid sequence ofVal⁸-Glu²²-GLP-1 (SEQ ID NO: 17) with two additional amino acid sequencesubstitutions. In another preferred embodiment, the combination of thepresent invention comprises an FGF-21 analog comprising the amino acidsequence of Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 10) in combinationwith a GLP-1 analog, wherein the GLP-1 analog comprises the amino acidsequence of Val⁸-Glu²²-GLP-1 (SEQ ID NO: 17) with one additional aminoacid sequence substitution. In another preferred embodiment, thecombination of the present invention comprises an FGF-21 analogconsisting of the amino acid sequence of Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21(SEQ ID NO: 10) in combination with a GLP-1 analog, wherein the GLP-1analog consists of the amino acid sequence of Val⁸-Glu²²-GLP-1 (SEQ IDNO: 17) with one additional amino acid sequence substitution.

Preferred FGF-21 compound and GLP-1 compound combinations of the presentinvention include, Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) incombination with Val⁸-GLP-1 (SEQ ID NO: 15),Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) in combination withVal⁸-Glu²²-GLP-1 (SEQ ID NO: 17). Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21(SEQ ID NO: 13) in combination with isVal⁸-Glu²²-Ile³³-Gly³⁶-Pro³⁷-Ser³⁸-Ser³⁹-Gly⁴¹-Ala⁴¹-Pro⁴²-Pro⁴³-Pro⁴⁴-Ser⁴⁵-GLP-1(SEQ ID NO: 19), Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) incombination with pegylated Val⁸-Glu²²-Ile³³-Cys-NH₂ ⁴⁶-GLP-1 (SEQ ID NO:21), Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) in combinationwith Arg³⁴-GLP-1 (SEQ ID NO: 16), Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷ FGF-21(SEQ ID NO: 13) in combination with acylated Arg³⁴-GLP-1 (SEQ ID NO:16), Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) in combinationwith Arg³⁴-Lys²⁶-(N-ε-(γ-Glu (N-α-hexadecanoyl)))-GLP-1 (SEQ ID NO: 18).Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) in combination withGly⁸-Gly²²-GLP-1-1L-IgG4 (S228P) (SEQ ID NO: 24),Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) withGly⁸-Glu²²-Gly³⁶-GLP-1-1L-IgG4 (S228P, F234A, L235A) (SEQ ID NO: 25),Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1-1 L-IgG4 (S228P, N297A) (SEQ ID NO: 26),Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1-1L-IgG4 (S228P, F234A. L235A, N297A) (SEQ ID NO:27), Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) in combinationwith Gly⁸-Glu²²-Gly³⁶-GLP-1-1 L-IgG4 (S228P, des K) (SEQ ID NO: 28),Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) withGly⁸-Glu²²-Gly³⁶-GLP-1-1L-IgG4 (S228P: F234A, L235A, des K) (SEQ ID NO:29), Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) in combinationwith Gly⁸-Glu²²-Gly³⁶-GLP-1-1L-IgG4 (S228P, N297A, des K) (SEQ ID NO:30), Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) in combinationwith Gly⁸-Glu²²-Gly³⁶-GLP-1-L-IgG4 (S228P, F234A, L235A, N297A, des K)(SEQ ID NO: 31), Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO:13) incombination with Gly⁸-Glu²²-Gly³⁶-GLP-1-1.5L-IgG4 (S228P) (SEQ ID NO:32), Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) in combinationwith Gly⁸-Glu²²-Gly³⁶-GLP-1-1.5L-IgG4 (S228P, F234A, L235A) (SEQ ID NO:33). Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) in combinationwith Gly⁸-Glu²²-Gly³⁶-GLP-1-1.5L-IgG4 (S228P, N297A) (SEQ ID NO: 34),Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1-1.5L-IgG4 (S228P, F234A, L235A, N297A) (SEQ IDNO: 35), Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) incombination with Gly⁸-Glu²²-Gly³⁶-GLP-1-1.5L-IgG4 (S228P, des K) (SEQ IDNO: 36), Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) incombination with Gly⁸-Glu²²-Gly³⁶-GLP-1-1.5L-IgG4 (S228P, F234A, L235A,des K) (SEQ ID NO: 37), Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷Ala¹⁶⁷-FGF-21 (SEQ IDNO: 13) in combination with Gly⁸-Glu²²-Gly³⁶-GLP-1-1.5L-IgG4 (S228P,N297A, des K) (SEQ ID NO: 38), Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQID NO: 13) in combination with Gly⁸-Glu²²-Gly³⁶-GLP-1-1.5L-IgG4 (S228P,F234A, L235A, N297A, des K) (SEQ ID NO: 39),Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1-2L-IgG4 (S228P) (SEQ ID NO: 40),Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1-2L-IgG4 (S228P, F234A, L235A) (SEQ ID NO: 41),Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1-2L-IgG4 (S228P, N297A) (SEQ ID NO: 42),Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1-2L-IgG4 (S228P, F234A, L235A, N297A) (SEQ ID NO:43), Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) in combinationwith Gly⁸-Glu²²-Gly³⁶-GLP-1-2L-IgG4 (S228P, des K) (SEQ ID NO: 44),Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) in combination withGly⁸-Glu²²-Gly³⁶-GLP-1-2L-IgG4 (S228P, F234A, L235A, des K) (SEQ ID NO:45), Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) in combinationwith Gly⁸-Glu²²-Gly³⁶-GLP-1-2L-IgG4 (S228P, N297A, des K) (SEQ ID NO:46). Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) in combinationwith Gly⁸-Glu²²-Gly³⁶-GLP-1-2L-IgG4 (S228P, F234A, L235A, N297A, des K)(SEQ ID NO: 47), Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) incombination with exendin-4 (SEQ ID NO: 4),Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) in combination withalbiglutide (SEQ ID NO: 49), Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO:10)in combination with a GLP-1 compound comprising the amino acid sequenceof SEQ ID NO: 47, Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO:10) incombination with a GLP-1 compound consisting of the amino acid sequenceof SEQ ID NO: 47, Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) incombination with a GLP-1 compound comprising the amino acid sequence ofSEQ ID NO: 48, Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) incombination with a GLP-1 compound consisting of the amino acid sequenceof SEQ ID NO: 48, Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) incombination with a GLP-1 compound comprising the amino acid sequence ofSEQ ID NO: 49, Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) incombination with a GLP-1 compound consisting of the amino acid sequenceof SEQ ID NO: 49, Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) incombination with a GLP-1 compound comprising the amino acid sequence ofSEQ ID NO: 50, Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) incombination with a GLP-1 compound consisting of the amino acid sequenceof SEQ ID NO: 50, Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) incombination with a GLP-1 compound comprising the amino acid sequence ofSEQ ID NO: 51, Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) incombination with a GLP-1 compound consisting of the amino acid sequenceof SEQ ID NO: 51, Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) incombination with a GLP-1 compound comprising the amino acid sequence ofSEQ ID NO: 52. Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) incombination with a GLP-1 compound consisting of the amino acid sequenceof SEQ ID NO: 52, Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) incombination with a GLP-1 compound comprising the amino acid sequence ofSEQ ID NO: 53, Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) incombination with a GLP-1 compound consisting of the amino acid sequenceof SEQ ID NO: 53, Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) incombination with a GLP-1 compound comprising the amino acid sequence ofSEQ ID NO: 54, Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) incombination with a GLP-1 compound consisting of the amino acid sequenceof SEQ ID NO: 54, Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) incombination with a GLP-1 compound comprising the amino acid sequence ofSEQ ID NO: 55, Cys¹¹⁸-Cys¹³⁴-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) incombination with a GLP-1 compound consisting of the amino acid sequenceof SEQ ID NO: 55, Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) incombination with a GLP-1 compound comprising the amino acid sequence ofSEQ ID NO: 56, Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) incombination with a GLP-1 compound consisting of the amino acid sequenceof SEQ ID NO: 56, Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) incombination with a GLP-1 compound comprising the amino acid sequence ofSEQ ID NO: 57, and Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) incombination with a GLP-1 compound consisting of the amino acid sequenceof SEQ ID NO: 57.

In a preferred embodiment, the combination of the present inventioncomprises an FGF-21 analog comprising the amino acid sequence ofCys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) in combination with aGLP-1 analog, wherein the GLP-1 analog comprises the amino acid sequenceof Val⁸-Glu²²-GLP-1 (SEQ ID NO: 17) with five additional amino acidsequence substitutions. In a preferred embodiment, the combination ofthe present invention comprises an FGF-21 analog consisting of the aminoacid sequence of Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) incombination with a GLP-1 analog, wherein the GLP-1 analog consists ofthe amino acid sequence of Val⁸-Glu²²-GLP-1 (SEQ ID NO: 17) with fiveadditional amino acid sequence substitutions. In another preferredembodiment, the combination of the present invention comprises an FGF-21analog comprising the amino acid sequence ofCys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) in combination with aGLP-1 analog, wherein the GLP-1 analog comprises the amino acid sequenceof Val⁸-Glu²²-GLP-1 (SEQ ID NO: 17) with four additional amino acidsequence substitutions. In another preferred embodiment, the combinationof the present invention comprises an FGF-21 analog consisting of theamino acid sequence of Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO:13) in combination with a GLP-1 analog, wherein the GLP-1 analogconsists of the amino acid sequence of Val⁸-Glu²²-GLP-1 (SEQ ID NO: 17)with four additional amino acid sequence substitutions. In anotherpreferred embodiment, the combination of the present invention comprisesan FGF-21 analog comprising the amino acid sequence ofCys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) in combination with aGLP-1 analog, wherein the GLP-1 analog comprises the amino acid sequenceof Val⁸-Glu²²-GLP-1 (SEQ ID NO: 17) with three additional amino acidsequence substitutions. In another preferred embodiment, the combinationof the present invention comprises an FGF-21 analog consisting of theamino acid sequence of Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO:13) in combination with a GLP-1 analog, wherein the GLP-1 analogconsists of the amino acid sequence of Val⁸-Glu²²-GLP-1 (SEQ ID NO: 17)with three additional amino acid sequence substitutions. In anotherpreferred embodiment, the combination of the present invention comprisesan FGF-21 analog comprising the amino acid sequence ofCys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) in combination with aGLP-1 analog, wherein the GLP-1 analog comprises the amino acid sequenceof Val⁸-Glu²²-GLP-1 (SEQ ID NO: 17) with two additional amino acidsequence substitutions. In another preferred embodiment, the combinationof the present invention comprises an FGF-21 analog consisting of theamino acid sequence of Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO:13) in combination with a GLP-1 analog, wherein the GLP-1 analogconsists of the amino acid sequence of Val⁸-Glu²²-GLP-1 (SEQ ID NO: 17)with two additional amino acid sequence substitutions. In anotherpreferred embodiment, the combination of the present invention comprisesan FGF-21 analog comprising the amino acid sequence ofCys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO: 13) in combination with aGLP-1 analog, wherein the GLP-1 analog comprises the amino acid sequenceof Val⁸-Glu²²-GLP-1 (SEQ ID NO: 17) comprises one additional amino acidsequence substitution. In another preferred embodiment, the combinationof the present invention comprises an FGF-21 analog consisting of theamino acid sequence of Cys¹¹⁸-Cys¹³⁴-Ala¹²¹-Ala¹⁶⁷-FGF-21 (SEQ ID NO:13) in combination with a GLP-1 analog, wherein the GLP-1 analogconsists of the amino acid sequence of Val⁸-Glu²²-GLP-1 (SEQ ID NO: 17)comprises one additional amino acid sequence substitution.

Example 1 Effect of Combination Treatment with GLP-1 and FGF-21 on BodyWeight and on Blood Glucose Levels

Diet-induced obese (DIO) male C57/Bl6 mice (Harlan: Virginia) maintainedon a calorie rich diet (TD95217, Teklad, Madison, Wis.) since weaningare used. DIO is established by ad libitum feeding for at least 7 weeksof a diet consisting of 40% fat, 39% carbohydrate, and 21% proteincaloric content (TD)95217). Animals are individually housed in atemperature-controlled (24° C.) facility with 12 hour light/dark cycle(lights on 2200) and free access to food (TD95217) and water. After aminimum of 2 weeks acclimation to the facility, the mice are randomizedaccording to their body weight, so each experimental group of animalswould have similar body weight.

Body composition of DIO male C57/B16 mice is determined by using QNMRanalysis 1 day prior to initiation of treatment. Combination treatmentis administered with two delivery methods. A FGF-21 compound (1 mg/kg)is subcutaneously injected once a day and a GLP-1 compound (3nmol/kg/day) is delivered by continuous subcutaneous infusion with alzetpump. The experimental groups are as follow: One group (“Vehicle”) ofmice (n=8) receives PBS (11.4 μL/day) and a daily subcutaneous injectionof 0.05 mL/10 g of PBS. Another group (“FGF-21”) of mice (n=8) receives11.4 μL/day of PBS using the alzet pump and a daily subcutaneousinjection of 1 mg/kg of FGF-21. The “GLP-1” group of mice receives 3nmol/kg/day of a GLP-1 compound through an alzet pump and a dailysubcutaneous injection of 0.05 mL/10 g of PBS. The “GLP-1+FGF-21” groupof mice receives 3 nmol/kg/day of a GLP-1 compound through an alzet pumpand a daily subcutaneous injection of 1 mg/kg of a FGF-21 compound.Injections occur prior to the onset of the dark photoperiod. Startingweight values are measured after implantation of alzet pumps on Day 1.Body weights are recorded in conjunction with daily dosing. Averageweight changes compared to starting weights of a particular treatmentgroup, including vehicle, are determined. Average daily weight changesare normalized to average of daily weight change of Vehicle group andare reported in Table 1 (i.e., (Average weight change of Treatmentgroup)−(Average weight change of Vehicle group)). For all treatmentgroups n=8.

Data in Table 1 represent results where the FGF-21 compound is humanFGF-21 (SEQ ID NO: 1) and where the GLP-1 compound isVal⁸-Glu²²-Ile³³-Gly³⁶-Pro³⁷-Ser³⁸-Ser³⁹-Gly⁴⁰-Ala⁴¹-Pro⁴²-Pro⁴³-Pro³⁴-Ser⁴⁵-GLP-1(SEQ ID NO: 19). The data demonstrate that the use of a FGF-21 compoundin combination with a GLP-1 compound results in a synergistic effect onweight loss.

TABLE 1 Average Weight Change Normalized to Vehicle Treated MiceTreatment FGF-21 GLP-1 GLP-1 + FGF-21 Day (g) (g) (g) Day 1 0 0 0 Day 5−0.45 −1.01 −2.62 Day 10 −1.94 −1.99 −5.16 Day 15 −2.85 −2.50 −6.94

1-15. (canceled)
 16. A method of treating diabetes comprisingadministering an FGF-21 compound in combination with a GLP-1 compound,wherein the GLP-1 compound comprises an amino acid sequence of SEQ IDNO: 20: His Xaa Glu Gly Thr Phe Thr Ser Asp Val Ser Ser           10                  15Tyr Leu Glu Xaa Gln Ala Ala Lys Glu Phe Ile Ala    20                  25                  30Trp Leu Xaa Lys Gly Gly Pro Ser Ser Gly Ala Pro                35                  40 Pro Pro Cys Xaa         45

wherein Xaa at position 8 is: D-Ala, Gly, Val, Leu, Ile, Ser, or Thr,Xaa at position 22 is: Gly, Glu, Asp, or Lys; Xaa at position 33 is: Valor Ile Xaa at position 46 is: Cys or Cys-NH₂ and wherein one PEGmolecule is covalently attached to Cys⁴⁵ and one PEG molecule iscovalently attached to Cys⁴⁶ or Cys⁴⁶-NH₂.
 17. A method of treatingdiabetes comprising administering an FGF-21 compound in combination witha GLP-1 compound, wherein the GLP-1 compound is a fusion proteincomprising a GLP-1 analog fused via a linker to an FC portion of animmunoglobulin, wherein the GLP-1 analog comprises an amino acidsequence of SEQ ID NO:22:His Xaa Glu Gly Thr Phe Thr Ser Asp Val Ser Ser            10                  15Tyr Leu Glu Glu Gln Ala Ala Lys Glu Phe Ile Ala    20                  25                  30Trp Leu Xaa Xaa Gly Gly Xaa                 35

wherein Xaa at position 8 is Gly or Val; Xaa at position 33 is Val orLys; Xaa at position 34 is Lys or Asn; Xaa at position 37 is Gly, Pro oris absent; the linker comprises an amino acid sequence of SEQ ID NO:6:Gly-Gly-Gly-Gly-Ser-Gly-Gly-Gly-Gly-Ser-Gly-Gly-Gly-Gly-Ser, and the Fcportion comprises an amino acid sequence of SEQ ID NO: 23:Ala Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys                5                   10Pro Ala Pro Xaa Xaa Xaa Gly Gly Pro Ser Val Phe        15                  20Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile25                  30                  35Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp            40                  45Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp    50                  55                  60Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr                65                  70Lys Pro Arg Glu Glu Gln Phe Xaa Ser Thr Tyr Arg        75                  80Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp85                  90                  95Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn            100                 105Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser    110                 115                 120Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr               125                  130Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn        135                 140Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr145                 150                 155Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly            160                 165Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val    170                 175                 180Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg                185                 190Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn        195                 200Val Phe Ser Cys Ser Val Met His Glu Ala Leu His205                 210                 215Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu            220                 225 Gly Xaa     230

wherein: Xaa at position 16 is Pro or Glu; Xaa at position 17 is Phe,Val, or Ala; Xaa at position 18 is Leu, Glu, or Ala; Xaa at position 80is Asn or Ala; and Xaa at position 230 is Lys or is absent.
 18. A methodof lowering blood glucose levels comprising administering an FGF-21compound in combination with a GLP-1 compound, wherein the GLP-1compound comprises an amino acid sequence of SEQ ID NO: 20:His Xaa Glu Gly Thr Phe Thr Ser Asp Val Ser Ser           10                  15Tyr Leu Glu Xaa Gln Ala Ala Lys Glu Phe Ile Ala    20                  25                  30Trp Leu Xaa Lys Gly Gly Pro Ser Ser Gly Ala Pro                35                  40 Pro Pro Cys Xaa         45

wherein Xaa at position 8 is: D-Ala, Gly, Val, Leu, Ile, Ser, or Thr,Xaa at position 22 is: Gly, Glu, Asp, or Lys; Xaa at position 33 is: Valor Ile Xaa at position 46 is: Cys or Cys-NH₂ and wherein one PEGmolecule is covalently attached to Cys⁴⁵ and one PEG molecule iscovalently attached to Cys⁴⁶ or Cys⁴⁶-NH₂.
 19. A method of loweringblood glucose levels comprising administering an FGF-21 compound incombination with a GLP-1 compound, wherein the GLP-1 compound is afusion protein comprising a GLP-1 analog fused via a linker to an FCportion of an immunoglobulin, wherein the GLP-1 analog comprises anamino acid sequence of SEQ ID NO: 22:His Xaa Glu Gly Thr Phe Thr Ser Asp Val Ser Ser            10                  15Tyr Leu Glu Glu Gln Ala Ala Lys Glu Phe Ile Ala    20                  25                  30Trp Leu Xaa Xaa Gly Gly Xaa                 35

wherein Xaa at position 8 is Gly or Val; Xaa at position 33 is Val orLys; Xaa at position 34 is Lys or Asn; Xaa at position 37 is Gly, Pro oris absent; the linker comprises an amino acid sequence of SEQ ID NO:6:Gly-Gly-Gly-Gly-Ser-Gly-Gly-Gly-Gly-Ser-Gly-Gly-Gly-Gly-Ser, and the Fcportion comprises an amino acid sequence of SEQ ID NO:23:Ala Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys                5                   10Pro Ala Pro Xaa Xaa Xaa Gly Gly Pro Ser Val Phe        15                  20Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile25                  30                  35Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp            40                  45 Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp    50                  55                  60Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr                65                  70Lys Pro Arg Glu Glu Gln Phe Xaa Ser Thr Tyr Arg        75                  80Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp85                  90                  95Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn            100                 105 Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser    110                 115                 120Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr               125                  130Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn        135                 140Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr145                 150                 155Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly            160                 165Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val    170                 175                 180Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg                185                 190Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn        195                 200Val Phe Ser Cys Ser Val Met His Glu Ala Leu His205                 210                 215Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu            220                 225 Gly Xaa     230

wherein: Xaa at position 16 is Pro or Glu; Xaa at position 17 is Phe,Val, or Ala; Xaa at position 18 is Leu, Glu, or Ala; Xaa at position 80is Asn or Ala; and Xaa at position 230 is Lys or is absent.
 20. Themethod according to claim 16, wherein the FGF21 compound comprises anamino acid sequence comprising a cysteine substitution at positions 21,26, 33, 118, 119, 121, 122, or 134 of FGF-21 (SEQ ID NO: 2).
 21. Themethod according to claim 17, wherein the FGF21 compound comprises anamino acid sequence comprising a cysteine substitution at positions 21,26, 33, 118, 119, 121, 122, or 134 of FGF-21 (SEQ ID NO: 2).
 22. Themethod according to claim 18, wherein the FGF21 compound comprises anamino acid sequence comprising a cysteine substitution at positions 21,26, 33, 118, 119, 121, 122, or 134 of FGF-21 (SEQ ID NO: 2).
 23. Themethod according to claim 19, wherein the FGF21 compound comprises anamino acid sequence comprising a cysteine substitution at positions 21,26, 33, 118, 119, 121, 122, or 134 of FGF-21 (SEQ ID NO: 2).